Studies On The Chemical Constituents And Biologicalactivities Of Aglaia Elliptifolia Merr.

Aglaia elliptifolia Merr.is an evergreen shrub or small tree of Meliaceae.Aglaia.and Rutales.In recent years,it has been reported that there are many kinds of compounds,such as diamide,tetracyclic triterpene,flavone and so on.Some of the compounds also have significant anti-inflammatory and anti-tumor effects.The study on the chemical components and physiological activities of Aglaia elliptifolia Merr.can not only develop and utilize the resources of Aglaia elliptifolia Merr.,but also provide scientific basis for some medical guidance.In this paper,20 monomer compounds extracted from the Aglaia elliptifolia Merr.were screened for antibacterial and antitumor activities in vitro.The mechanism of compound 11 on HCT-116 cells was studied.Objective: In this paper,the structure of Aglaia elliptifolia Merr.was identified by various chromatographic separation methods and modern spectral analysis techniques,and the monomer compounds obtained enriched the compound library of Aglaia elliptifolia Merr..At the same time,the new active components in Aglaia elliptifolia Merr.were searched through the biological activity screening of 20 monomer compounds,which provided for the further development and utilization of Aglaia elliptifolia Merr.certain scientific basis.Methods:(1)Through silica gel column chromatography,ODS column chromatography an d semi preparative HPLC and other methods for separation and purification,through 1H-NMR,13 CNMR,ESI-MS and other methods to analyze the structure of the compounds,determine the structu re of the monomer of Aglaia elliptifolia Merr.(2)Study on the effect of 20 monomers of Aglaia elliptifolia Merr.on Staphylococcus aureus(3)Study on the antitumor activity of 20 monomers of Aglaia elliptifolia Merr.in vitro: A549(human lung adenocarcinoma cell line),HL-60(human pr omyelocytic leukemia cell line),HT-29(Human colon cancer cell line),HCT-116(Human colorecta l cancer cell line),MDA-MB-231(Human breast cancer cell line),K562(Human chronic myeloid leukemia cell line),and other tumor cells.At the same time,the selective effect of HL-7702(nor mal human liver cell line)was investigated.Compound 11 showed the best activity.Therefore,the effect of compound 11 on autophagy was further analyzed by flow cytometry with MDC stainingafter drug treatment.Immunofluorescence of LC3 B was also applied to detect autophagy,Western Blot was used to detect the changes of protein expression like LC3B?P62?PINK1/Parkin?LAMP1?TOMM20 after compound 11 treatment on HCT-116 cells.Results:(1)Twenty compounds were isolated from Aglaia elliptifolia Merr.by the above method,These 20 monomers were isolated from this genus for the first time.They were identified as: Daucosterol(1),sitosterol-3-O-6-palmityl-?-D-glucopyranoside(2),(+)-catechin(3),Epicatechin(4),Lyratol F(5),3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone(6),?-sitosterol(7),Yohimbe(8),?-Tripalmiti(9),4-(aminomethyl)-2-benzyl-5-(ethylamino)pyridazin-3(2H)-one(10),Epigallocatechin(11),gallocatechin-(4'-O-7)-epigallocatechin(12),(-)-lyoniresinol 3?-O-?-D-glucopyranoside(13),(+)-is olariciresinol-9'-O-?-glucopyranosidede(14),(6S,7S,8R)-7?-[?-D-glucopyranosyl)oxy]isolariciresinol(15),procyanidin B3(16),(2R,3S)-5,7,3'-tri-Omethyl-(-)-epicatechin(17),procyanidin B1(18),procyanidin B2(19),(±)-6-linalyl-7-hydroxycoumarin(20).(2)The compounds 20 extracted from Aglaia elliptifoli a Merr.were screened for the activity against Staphylococcus aureus.The experimental results sho wed that compound 4,11,12,16,18,19 exhibited antibacterial activity.The minimum inhibitory concen tration(MIC)were 1.0 mg/m L,1.0 mg/m L,1.0 mg/m L,1.0 mg/m L,1.0 mg/m L,0.8 mg/m L,respecti vely.(3)MTT method was used to detect the inhibitory effect of 20 monomers extracted from Ag laia elliptifolia Merr.on the proliferation of tumor cells.The results showed that 6,11,12,13,18,19,20 compounds had inhibitory effects on cell proliferation,which showed different cytotoxicity to sev en cell lines.(4)The results showed that compound 11 had the best antitumor activity on HCT-116 cells.Therefore,compound 11 was further analyzed by flow cytometry for MDC staining.The r esults showed that compound 11 could increase the autophagy rate in a dose-dependent manner,w hich was statistically significant compared with the control group(P<0.05).The results of LC3 B st aining further showed that compound 11 could significantly increase the positive rate of LC3 B in HCT-116 cells,suggesting that autophagy increased in a concentration dependent manner,which wa s statistically significant compared with the control group(P<0.05).Western Blot wasused to detect the effect of autophagy-related protein by compound 11 treatment.the protein level of LC3B?PI NK1/Parkin?LAMP1 was elevated and the expression of p62?TOMM20 was decreased.It suggest edcompound 11 may active PINK1/Parkin mediated mitophagy and lead to cell death in HCT-116 cells.